Automated Focal Plane Merging From a Stack of Gonioscopic Photographs Using a Focus-Stacking Algorithm.

Purpose: The purpose of this study was to investigate the utility of automated focal plane merging with the collection of gonio-photographs with different depths of field (DOF) using an established focus-stacking algorithm. Methods: A cross-sectional study was conducted at Shimane University Hospital, Izumo, Japan. Sixteen eyes from 16 subjects from the glaucoma clinic were included in this study. Image processing was performed for the images of 16 eyes from 16 angle sector following the successful gonio-photography. The 256 sets of focus-stacked and best-focused images were prepared in random order and were compared for the DOF and informativeness to diagnose angle pathology by masked observers in each set as the subjective assessments. Moreover, the energy of the Laplacian (average |ΔI|), which is an indicator of image sharpness between the photographs with and without the focus-stacking processing was also analyzed with the Laplacian filter as the objective assessment. Results: The automated image processing was successfully performed in all stacks of images. The significant deepening of DOF and improvement of informativeness achieved in 255 (99.6%) and 216 (84.4%) images (P < 0.0001 for both, sign test) and the energy of the Laplacian also significantly increased in 243 (94.9%) images (P < 0.0001, sign test). Conclusions: Focal plane merging by the automated algorithm can make the gonio-images deeper focus compared with the paired best-focused images subjectively and objectively, which would be useful for angle pathological assessment in clinical practice. Translational Relevance: Focal plane merging algorithm for the automated gonio-photography can facilitate the angle assessment by providing informative deep-focus image, which would be useful for glaucoma care.

Anti-vascular Endothelial Growth Factor Antibody Limits the Vascular Leakage and Decreases Subretinal Fibrosis in a Cynomolgus Monkey Choroidal Neovascularization Model.

OBJECTIVE: This study was conducted to evaluate the effects of anti-vascular endothelial growth factor (VEGF) antibody (bevacizumab) on vascular leakage and fibrosis in a monkey choroidal neovascularization (CNV) model. The relationship between fibrotic tissue and subretinal hyper-reflective material (SHRM), in optical coherence tomography (OCT) images, was also investigated. METHODS: Experimental CNV was induced in male cynomolgus monkeys by laser photocoagulation. Intravitreal injection of bevacizumab at 0.5 mg/eye/dosing was initiated 2 weeks before or after laser irradiation and thereafter, conducted intermittently at 2- or 3-week intervals. Fluorescein fundus angiography (FA) and OCT imaging were conducted weekly from 2 to 7 weeks after laser irradiation. CNV leakage was evaluated by an established grading method using FA images. To assess the fibrosis and scarring, Masson's trichrome specimens of each CNV lesion were prepared, and morphometric analysis was conducted using an image analysis software. RESULTS: The effects of bevacizumab on vascular leakage were shown using an established evaluation method. Morphometric analysis of Masson's trichrome-stained (MT) specimens revealed that collagen fiber synthesis was suppressed by bevacizumab pre-treatment (-29.2%) or post-treatment (-19.2%). SHRM was detected in OCT images in a monkey CNV model, and a significant correlation between the SHRM area in the OCT images and the collagen fiber area in the MT specimens was noted. CONCLUSION: In the established cynomolgus monkey CNV model, bevacizumab prevented blood leakage but could not completely suppress fibrosis. SHRM in the OCT images reflected retinal fibrous tissue in a laser-induced CNV monkey model. This model might be useful for elucidating the pathology and development therapy for neovascularization or fibrosis.

Relationship between Elevated Intraocular Pressure and Divided Peripapillary Sector Retinal Nerve Fiber Layer Thickness in a Cynomolgus Monkey Laser-Induced Ocular Hypertension Model.

AIMS: We investigated the relationship between elevated intraocular pressure (IOP) and changes in global and peripapillary sector retinal nerve fiber layer (RNFL) thickness around the optic nerve head (ONH) in the laser-induced ocular hypertension monkey model. METHODS: To induce high IOP, green laser photocoagulation burns were applied around the trabecular meshwork of 1 eye from each of 12 cynomolgus monkeys. The animals had been acclimated to IOP measurement under conscious conditions for more than 2 months, and IOP was chronologically measured. RNFL thickness was measured for 6 peripapillary sectors and global area using spectral-domain optical coherence tomography. RESULTS: After model induction, marked IOP elevation and enlarged optic disk cupping were observed. Thinning of the RNFL associated with elevated IOP was observed around the ONH from 6 until 9 weeks after laser treatment, and the degree of reduction in RNFL thickness varied between the peripapillary sectors. Correlations between cumulative IOP elevation and RNFL thickness reduction were statistically significant for the temporal-superior (p = 0.024), nasal-inferior (p = 0.044), and temporal (p = 0.049) sectors, and global RNFL (p = 0.018). CONCLUSIONS: These results suggest that this model reflected the pathology of clinical glaucoma in terms of the specific pattern of RNFL thinning around the ONH.